Nod-factor perception
The present invention provides a Nod-factor binding element, comprising one or more NFR polypeptides encoded by NFR genes, that are useful for providing non-nodulating plants with Nod-factor binding properties and triggering the endosymbiotic signalling pathway leading to nodulation. Furthermore the invention is useful for breeding for improved nodulation in nodulating legumes.
1. An isolated Nod-factor binding polypeptide comprising:
at least 80% amino acid sequence identity to any one of SEQ ID NO: 8, 15, 31, 32, 40, or 48, wherein said polypeptide comprises an extracellular domain comprising 2 or 3 different LysM-type motifs, and wherein said polypeptide selectively binds strain-specific forms of Nod-Factor.
2. An isolated Nod-factor binding polypeptide comprising: at least 80% amino acid sequence identity to any one of SEQ ID NO: 24 or 25, wherein said polypeptide comprises an extracellular domain comprising 2 or 3 different LysM-type motifs, and wherein said polypeptide selectively binds strain-specific forms of Nod-Factor.
3. A method of producing a transgenic plant expressing a Nod-factor binding polypeptide, the method comprising:
a. introducing into the plant a nucleic acid molecule encoding one or more Nod- factor binding polypeptide of claim 2 , wherein the nucleic acid molecule is operably linked to a promoter; and
b. selecting transgenic plants expressing the Nod-factor binding polypeptide.
4. The isolated Nod-factor binding polypeptide of claim 2 , wherein said polypeptide comprises the amino acid sequence of any one of SEQ ID NO: 24 or 25.
5. An isolated Nod-factor binding element comprising one or more isolated Nod-factor binding polypeptide of claim 1 , and further comprising one or more isolated Nod-factor binding polypeptide comprising at least 80% amino acid sequence identity to any one of SEQ ID NO: 24, 25, 52, or 54, wherein said polypeptide comprises an extracellular domain comprising 2 or 3 different LysM-type motifs, and wherein said polypeptide selectively binds strain-specific forms of Nod-Factor.
6. An isolated Nod-factor binding element comprising one or more isolated Nod-factor binding polypeptide of claim 3 , and further comprising one or more polypeptide comprising the amino acid sequence of any one of SEQ ID NO: 24, 25, 52, or 54.
7. An isolated nucleic acid molecule encoding the Nod-factor binding polypeptide of claim 1 .
8. An isolated nucleic acid molecule encoding the Nod-factor binding polypeptide of claim 2 .
9. The isolated nucleic acid molecule of claim 7 , wherein said nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 6, 7, 11, 12, 30, 39, or 47.
10. The isolated nucleic acid molecule of claim 8 , wherein said nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 21, 22, or 23.
11. A transgenic cell stably transformed with one or more nucleic acid molecule encoding the Nod-factor binding polypeptide of claim 1 .
12. The transgenic cell of claim 11 , wherein said nucleic acid molecule encodes a polypeptide having the sequence of SEQ ID NOS: 8, 15, 31, 32, 40, or 48.
13. The transgenic cell of claim 11 , wherein said nucleic acid molecule comprises the sequence of SEQ ID NOS: 6, 7, 11, 12, 30, 39, or 47.
14. A transgenic cell stably transformed with one or more nucleic acid molecule encoding the Nod-factor binding polypeptide of claim 2 .
15. The transgenic cell of claim 14 , wherein said nucleic acid molecule encodes a polypeptide having the sequence of SEQ ID NOS: 24 or 25.
16. The transgenic cell of claim 14 , wherein said nucleic acid molecule comprises the sequence of SEQ ID NOS: 21, 22, or 23.
17. A transgenic cell comprising one or more transgene encoding the Nod Factor binding element of claim 5 .
18. A transgenic cell comprising one or more transgene encoding the Nod Factor binding element of claim 6 .
19. A method of producing a transgenic plant expressing a Nod-factor binding polypeptide, the method comprising:
a. introducing into the plant a nucleic acid molecule encoding one or more Nod-factor binding polypeptide of claim 1 , wherein the nucleic acid molecule is operably linked to a promoter; and
b. selecting transgenic plants expressing the Nod-factor binding polypeptide.
20. The method of claim 19 , wherein said nucleic acid molecule encodes a polypeptide having the amino acid sequence of SEQ ID NO: 8, 15, 31, 32, 40, or 48.
21. The method of claim 19 , wherein said nucleic acid molecule comprises the sequence of SEQ ID NO: 6, 7, 11, 12, 30, 39, or 47.
22. A method of producing a transgenic plant expressing a Nod-factor binding polypeptide, the method comprising:
a. introducing into the plant a nucleic acid molecule encoding one or more Nod-factor binding polypeptide of claim 2 , wherein the nucleic acid sequence is operably linked to a promoter; and
b. selecting transgenic plants expressing the Nod-factor binding polypeptide.
23. The method of claim 22 , wherein said nucleic acid molecule encodes a polypeptide having the amino acid sequence of SEQ ID NO: 24 or 25.
24. The method of claim 22 , wherein said nucleic acid molecule comprises the sequence of SEQ ID NO: 21, 22, or 23.
25. The method of claim 19 , further comprising introducing into the plant one or more nucleic acid molecule encoding a polypeptide having at least 80% amino acid sequence identity to SEQ ID NO: 24, 25, 52, or 54.
26. The method of claim 20 , comprising:
introducing into the plant one or more nucleic acid molecule encoding a polypeptide having the amino acid sequence of SEQ ID NO: 8, 15, 31, 32, 40, or 48; and further introducing into the plant one or more nucleic acid molecule encoding a polypeptide having the amino acid sequence of SEQ ID NO: 24, 25, 52, or 54.
27. The method of claim 25 , comprising introducing into the plant one or more nucleic acid molecule comprising SEQ ID NO: 6, 7, 11, 12, 30, 39, or 47; and further introducing one or more nucleic acid molecule comprising SEQ ID NO: 21, 22, 23, 51, or 53.
28. The method of claim 19 , wherein one or more nucleic acid molecule is introduced into the plant through a sexual cross.
29. The method of claim 22 , wherein one or more nucleic acid molecule is introduced into the plant through a sexual cross.
30. The method of claim 25 , wherein one or more nucleic acid molecule is introduced into the plant through a sexual cross.
31. The method of claim 27 , wherein one or more nucleic acid molecule is introduced into the plant through a sexual cross.
32. A transgenic plant comprising one or more transgene encoding the Nod-factor binding polypeptide of claim 1 .
33. The transgenic plant of claim 32 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 8, 15, 31, 32, 40, or 48.
34. A transgenic plant comprising one or more transgene encoding the Nod-factor binding polypeptide of claim 2 .
35. The transgenic plant of claim 34 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 24 or 25.
36. A transgenic plant comprising one or more transgene encoding the Nod-factor binding element of claim 5 .
37. A transgenic plant comprising one or more transgene encoding the Nod-factor binding element of claim 6 .
38. The transgenic plant of claim 32 , wherein said plant is a cereal.
39. The transgenic plant of claim 34 , wherein said plant is a cereal.
40. The transgenic plant of claim 32 , wherein said plant is a legume.
41. The transgenic plant of claim 34 , wherein said plant is a legume.
42. The transgenic plant of claim 32 , wherein said plant is a non-nodulating plant.
43. The transgenic plant of claim 34 , wherein said plant is a non-nodulating plant.
44. An isolated Nod-factor binding polypeptide comprising:
at least 90% amino acid sequence identity to SEQ ID NO: 52 or 54, wherein said polypeptide comprises an extracellular domain comprising 2 or 3 different LysM-type motifs, and wherein said polypeptide selectively binds strain-specific forms of Nod-Factor.
45. An isolated nucleic acid molecule encoding the Nod-factor binding polypeptide of claim 44 .
46. A transgenic cell stably transformed with one or more nucleic acid molecule encoding the Nod-factor binding polypeptide of claim 44 .
47. The transgenic cell of claim 46 , wherein said nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 51 or 53.
48. A transgenic plant comprising one or more transgene encoding the Nod-factor binding polypeptide of claim 44 .
49. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising at least 80% amino acid sequence identity to SEQ ID NO: 8.
50. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising at least 80% amino acid sequence identity to SEQ ID NO: 15.
51. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising at least 80% amino acid sequence identity to SEQ ID NO: 31.
52. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising at least 80% amino acid sequence identity to SEQ ID NO: 32.
53. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising at least 80% amino acid sequence identity to SEQ ID NO: 40.
54. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising at least 80% amino acid sequence identity to SEQ ID NO: 48.
55. The transgenic plant of claim 34 , wherein the transgene encodes a polypeptide comprising at least 80% amino acid sequence identity to SEQ ID NO: 24.
56. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising the sequence of SEQ ID NO: 8.
57. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising the sequence of SEQ ID NO: 15.
58. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising the sequence of SEQ ID NO: 31.
59. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising the sequence of SEQ ID NO: 32.
60. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising the sequence of SEQ ID NO: 40.
61. The transgenic plant of claim 32 , wherein the transgene encodes a polypeptide comprising the sequence of SEQ ID NO: 48.
62. The transgenic plant of claim 34 , wherein the transgene encodes a polypeptide comprising the sequence of SEQ ID NO: 24.
63. The transgenic plant of claim 48 , wherein the transgene encodes a polypeptide comprising the sequence of SEQ ID NO: 52.